TNF-α通过调节神经酰胺水平调控人乳腺癌细胞的增殖
发布时间：2018-11-11 07:56
【摘要】：目的检测肿瘤坏死因子α(TNF-α)对MCF-7和MDA-MB-231人乳腺癌细胞增殖的影响及神经酰胺(Cer)的介导作用。方法首先确定TNF-α的最佳处理剂量和时间,免疫荧光细胞化学染色结合共聚焦显微镜技术检测细胞Cer含量的变化,采用50 ng/mL TNF-α单独或联合ASM抑制剂地昔帕明(Des)、神经鞘氨醇激酶1(SPHK1)抑制剂N,N,-二甲基鞘氨醇(DMS)处理乳腺癌细胞,检测其在TNF-α所致细胞增殖变化中的作用。采用MTT法检测细胞增殖;Western blot法检测酸性鞘磷脂酶(ASM)、SPHK1及凋亡与增殖相关蛋白Bcl2、Bax、增殖细胞核抗原(PCNA)表达。结果 TNF-α可抑制MCF-7细胞增殖,促进MDA-MB-231细胞增殖,且具剂量和时间依赖性;在MCF-7细胞,TNF-α可显著增加ASM蛋白表达,对SPHK1蛋白表达无明显影响,而在MDA-MB-231细胞,可同时增加ASM和SPHK1蛋白表达;TNF-α作用后,MCF-7细胞Cer含量显著增加,ASM抑制剂Des预处理可抑制此作用;TNF-α作用可降低MDA-MB-231细胞Cer含量,此作用可被SPHK1抑制剂DMS所阻止;Des可抑制TNF-α作用所导致的MCF-7细胞增殖降低、Bcl2表达降低、Bax表达增加;DMS可抑制TNF-α作用所导致的MDA-MB-231细胞PCNA和细胞增殖指数的高表达。结论 TNF-α通过调节ASM和SPHK1蛋白的表达水平调控Cer生成,影响MCF-7和MDA-MB-231人乳腺癌细胞的增殖。
[Abstract]:Objective to investigate the effect of tumor necrosis factor 伪 (TNF- 伪) on the proliferation of MCF-7 and MDA-MB-231 human breast cancer cells and the mediated effect of ceramide (Cer). Methods the optimal treatment dose and time of TNF- 伪 were first determined. The changes of Cer content in cells were detected by immunofluorescence cytochemical staining combined with confocal microscopy. 50 ng/mL TNF- 伪 was used alone or in combination with ASM inhibitor desipramine (Des),. Breast cancer cells were treated with neurinosinolkinase-1 (SPHK1) inhibitor, NNN-dimethyl-sphingosine (DMS), and their role in the proliferation of breast cancer cells induced by TNF- 伪 was examined. The expression of Bcl2,Bax, proliferating cell nuclear antigen (PCNA) and apoptosis and sphingomyelinase (ASM), SPHK1) were detected by MTT assay by; Western blot assay. Results TNF- 伪 inhibited the proliferation of MCF-7 cells and promoted the proliferation of MDA-MB-231 cells in a dose-and time-dependent manner. In MCF-7 cells, TNF- 伪 could significantly increase the expression of ASM protein, but had no effect on the expression of SPHK1 protein, while in MDA-MB-231 cells, it could increase the expression of ASM and SPHK1 simultaneously. After the treatment of TNF- 伪, the content of Cer in MCF-7 cells increased significantly, which was inhibited by Des pretreatment with ASM inhibitor Des, and the Cer content of MDA-MB-231 cells was decreased by TNF- 伪, which was blocked by DMS, a SPHK1 inhibitor. Des inhibited the proliferation of MCF-7 cells induced by TNF- 伪, decreased the expression of Bcl2 and increased the expression of Bax, and DMS inhibited the high expression of PCNA and proliferation index of MDA-MB-231 cells induced by TNF- 伪. Conclusion TNF- 伪 affects the proliferation of MCF-7 and MDA-MB-231 human breast cancer cells by regulating the expression of ASM and SPHK1 protein.
【作者单位】： 空军军医大学西京医院甲乳血管外科;
【基金】：国家自然科学基金面上项目(81470120,81672593)
【分类号】：R737.9
[Abstract]:Objective to investigate the effect of tumor necrosis factor 伪 (TNF- 伪) on the proliferation of MCF-7 and MDA-MB-231 human breast cancer cells and the mediated effect of ceramide (Cer). Methods the optimal treatment dose and time of TNF- 伪 were first determined. The changes of Cer content in cells were detected by immunofluorescence cytochemical staining combined with confocal microscopy. 50 ng/mL TNF- 伪 was used alone or in combination with ASM inhibitor desipramine (Des),. Breast cancer cells were treated with neurinosinolkinase-1 (SPHK1) inhibitor, NNN-dimethyl-sphingosine (DMS), and their role in the proliferation of breast cancer cells induced by TNF- 伪 was examined. The expression of Bcl2,Bax, proliferating cell nuclear antigen (PCNA) and apoptosis and sphingomyelinase (ASM), SPHK1) were detected by MTT assay by; Western blot assay. Results TNF- 伪 inhibited the proliferation of MCF-7 cells and promoted the proliferation of MDA-MB-231 cells in a dose-and time-dependent manner. In MCF-7 cells, TNF- 伪 could significantly increase the expression of ASM protein, but had no effect on the expression of SPHK1 protein, while in MDA-MB-231 cells, it could increase the expression of ASM and SPHK1 simultaneously. After the treatment of TNF- 伪, the content of Cer in MCF-7 cells increased significantly, which was inhibited by Des pretreatment with ASM inhibitor Des, and the Cer content of MDA-MB-231 cells was decreased by TNF- 伪, which was blocked by DMS, a SPHK1 inhibitor. Des inhibited the proliferation of MCF-7 cells induced by TNF- 伪, decreased the expression of Bcl2 and increased the expression of Bax, and DMS inhibited the high expression of PCNA and proliferation index of MDA-MB-231 cells induced by TNF- 伪. Conclusion TNF- 伪 affects the proliferation of MCF-7 and MDA-MB-231 human breast cancer cells by regulating the expression of ASM and SPHK1 protein.
【作者单位】： 空军军医大学西京医院甲乳血管外科;
【基金】：国家自然科学基金面上项目(81470120,81672593)
【分类号】：R737.9
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